Subgenotypes and mutations in the s and polymerase genes of hepatitis B virus carriers in the West Bank, palestine.

The mutation rate and genetic variability of hepatitis B virus (HBV) are crucial factors for efficient treatment and successful vaccination against HBV. Until today, genetic properties of this virus among the Palestinian population remain unknown. Therefore, we performed genetic analysis of the overlapping S and polymerase genes of HBV, isolated from 40 Palestinian patients' sera. All patients were HBsAg positive and presented with a viral load above 105 HBV genome copies/ml. The genotyping results of the S gene demonstrated that HBV D1 was detected in 90% of the samples representing the most prominent subgenotype among Palestinians carrying HBV. Various mutations existed within the S gene; in five patients four known escape mutations including the common G145R and D144E were found. Furthermore, a ratio of 4.25 of non-synonymous to synonymous mutations in the S gene indicated a strong selection pressure on the HBs antigen loops of HBV strains circulating in those Palestinian patients. Although all patients were treatment-naïve, with the exception of one, several mutations were found in the HBV polymerase gene, but none pointed to drug resistance. The study presented here is the first report to address subgenotypes and mutation analyses of HBV S and polymerase genes in Palestine.

Seroprevalence of Cytomegalovirus among pregnant women and hospitalized children in Palestine.

BACKGROUND: Human Cytomegalovirus (HCMV) is the most common cause of congenital infections. The maternal immune status plays a major role in the likelihood of congenital infection. The aim of this study is to shed light on the seroprevalence of HCMV in pregnant women, hospitalized children and newborns including cases of congenital infections in Palestine. METHODS: We analyzed HCMV IgG and IgM test results that had been ordered for pregnant women, hospitalized children and newborns in the years 2006-2012 at Al-Makassed Islamic Charitable Hospital (MICH) in East Jerusalem. Furthermore, we reviewed the medical charts of newborns and HCMV IgM-positive children. RESULTS: HCMV IgG was positive in 96.6% of pregnant women, in 88% of hospitalized children and in 98.4% of hospitalized newborns. HCMV IgM was positive in 11.5% of pregnant women, in 11.7% of hospitalized children and in 2% of hospitalized newborns respectively. The HCMV avidity assay revealed that 95% of IgM-positive pregnant women had high avidity (>60%) indicating that most Palestinian women were undergoing a recurrent HCMV infection. Real time PCR on limited number of cases indicated that 62.5% of infants, mostly born to IgM-positive mothers and 83.3% of HCMV IgM-positive children had detectable HCMV DNA in their urine. Out of the 249 newborns tested during this study period, four (1.6%) were subjected to Gancyclovir treatment because of symptomatic congenital HCMV infection. CONCLUSIONS: This is the first report to provide an insight into HCMV seroprevalence in Palestine. Despite the high rate of seropositivity, the importance of HCMV testing during pregnancy should not be underestimated. A comprehensive study with a long term follow-up examination of offspring born to HCMV IgM-positive mothers would be required to provide estimates of an accurate percentage of symptomatic congenital HCMV infection in Palestine.

Analysis of human cytomegalovirus-encoded microRNA activity during infection.

MicroRNAs (miRNAs) are expressed in a wide variety of organisms, ranging from plants to animals, and are key posttranscriptional regulators of gene expression. Virally encoded miRNAs are unique in that they could potentially target both viral and host genes. Indeed, we have previously demonstrated that a human cytomegalovirus (HCMV)-encoded miRNA, miR-UL112, downregulates the expression of a host immune gene, MICB. Remarkably, it was shown that the same miRNA also downregulates immediate-early viral genes and that its ectopic expression resulted in reduced viral replication and viral titers. The targets for most of the viral miRNAs, and hence their functions, are still unknown. Here we demonstrate that miR-UL112 also targets the UL114 gene, and we present evidence that the reduction of UL114 by miR-UL112 reduces its activity as uracil DNA glycosylase but only minimally affects virus growth. In addition, we show that two additional HCMV-encoded miRNAs, miR-US25-1 and miR-US25-2, reduce the viral replication and DNA synthesis not only of HCMV but also of other viruses, suggesting that these two miRNAs target cellular genes that are essential for virus growth. Thus, we suggest that in addition to miR-UL112, two additional HCMV miRNAs control the life cycle of the virus.

Artesunate as a potent antiviral agent in a patient with late drug-resistant cytomegalovirus infection after hematopoietic stem cell transplantation.

This is the first report of treatment of cytomegalovirus infection with artesunate, for a stem cell transplant recipient with a newly identified foscarnet-resistant and ganciclovir-resistant DNA polymerase L776M mutation. Artesunate treatment resulted in a 1.7-2.1-log reduction in viral load by treatment day 7, with a viral half-life of 0.9-1.9 days, indicating a highly effective block in viral replication.

Comparison of human metapneumovirus, respiratory syncytial virus and influenza A virus lower respiratory tract infections in hospitalized young children.

BACKGROUND: We compared the clinical and demographic features of children with lower respiratory tract infection (LRI) caused by human metapneumovirus (HMPV), respiratory syncytial virus (RSV) and influenza A virus and sought to determine whether coinfection by HMPV and other respiratory viruses leads to increased disease severity. METHODS: Nasal wash specimens were prospectively obtained from 516 children hospitalized for LRI during a 1-year period and tested for the presence of HMPV by reverse transcription-polymerase chain reaction and for RSV and influenza A by direct immunofluorescence. RESULTS: HMPV was detected in 68 (13%) patients and was the third most common viral pathogen; 16 of 68 HMPV-positive children (24%) had coinfection with other respiratory viruses (HMPVco).HMPV patients were older than RSV patients (17.6 +/- 16.8 months versus 10.5 +/- 11.8 months, P = 0.02). HMPV was associated with wheezing and hypoxemia at a rate similar to that of RSV and higher than that of influenza A. Atelectasis was more common among HMPV (40%) than among RSV and influenza patients (13%, P < 0.05 for each). HMPV infection was more often associated with a diagnosis of pneumonia than RSV and influenza A and was more often associated with a diagnosis of asthma and less often associated with a diagnosis of bronchiolitis than RSV infection (P < 0.05 for each), even when corrected for age. Children with HMPVco had a higher rate of gastrointestinal symptoms but did not show a more severe respiratory picture. CONCLUSIONS: The clinical pattern of HMPV more closely resembles that of RSV than that of influenza A LRI, yet the differences in age, radiographic findings and clinical diagnosis suggest that HMPV pathogenesis may differ from that of RSV.

Early-onset Guillain-Barré syndrome associated with reactivation of Epstein-Barr virus infection after nonmyeloablative stem cell transplantation.

We report a case of early-onset acute Guillain-Barre syndrome associated with reactivation of Epstein-Barr virus (EBV) infection after nonmyeloablative stem cell transplantation (NST). Reactivation of EBV infection preceded disease onset, and the virus load increased concomitantly with disease progression (doubling time, 2.7 days). This case raises concern about the expanding scope of manifestations associated with reactivation of EBV infection after NST.

Emergence of late cytomegalovirus central nervous system disease in hematopoietic stem cell transplant recipients.

Preemptive ganciclovir therapy has reduced the occurrence of early cytomegalovirus (CMV) disease after hematopoietic stem cell (HSC) transplantation. However, late disease is increasingly reported. We describe 2 patients who developed late CMV central nervous system (CNS) disease after haploidentical HSC transplantation. Direct genotypic analysis was used to examine the presence of ganciclovir resistance. One patient had a mixed viral population in the cerebrospinal fluid (CSF), with coexistent wild-type and mutant UL97 sequences. The presence of 2 different strains was confirmed by subclone sequencing of the UL54 gene. One of the strains was different from the concurrent blood strain. The second patient had resistant variant in the lungs. These cases raise concern about the changing natural history of CMV disease in HSC transplantation, with emergence of previously uncommon manifestations following prolonged prophylaxis. Under these circumstances the CNS may be a sanctuary site, where viral persistence and antiviral drug resistance could result from limited drug penetration.